Long-term safety and efficacy of bimatoprost solution 0·03% application to the eyelid margin for the treatment of idiopathic and chemotherapy-induced eyelash hypotrichosis: a randomized controlled trial.

BACKGROUND: Bimatoprost ophthalmic solution 0·03% is approved in several countries for the treatment of eyelash hypotrichosis. Previous trials were limited to 4 months of treatment and primarily idiopathic hypotrichosis. OBJECTIVES: To evaluate the long-term safety and efficacy of bimatoprost in patients with idiopathic or chemotherapy-induced hypotrichosis. METHODS: This multicentre, double-masked, randomized, parallel-group study included two 6-month treatment periods [treatment period 1 (TP1) and treatment period 2 (TP2)]. Patients with idiopathic hypotrichosis were randomized to three treatment groups: (i) bimatoprost (TP1 and TP2); (ii) bimatoprost (TP1) and vehicle (TP2); and (iii) vehicle (TP1) and bimatoprost (TP2). Patients with chemotherapy-induced hypotrichosis were randomized to two treatment groups: (i) bimatoprost or vehicle (TP1) and (ii) bimatoprost (TP2). Primary end point was a composite of at least a one-grade improvement in investigator-assessed Global Eyelash Assessment and at least a three-point improvement in patient-reported Eyelash Satisfaction Questionnaire Domain 2 at month 4. Secondary measures included digitally assessed eyelash characteristics. RESULTS: The primary efficacy end point was met in both populations (idiopathic responder rate was 40·2% for bimatoprost vs. 6·8% for vehicle; postchemotherapy responder rate was 37·5% for bimatoprost vs. 18·2% for vehicle). Efficacy by month 6 was maintained (idiopathic) or enhanced (postchemotherapy) at 12 months. Treatment effects were maintained for approximately 2 months but markedly diminished 4-6 months following treatment cessation in patients with idiopathic hypotrichosis. No drug-related serious adverse events were reported. CONCLUSIONS: Daily treatment with bimatoprost ophthalmic solution 0·03% for 1 year was effective and well tolerated in patients with idiopathic and chemotherapy-induced hypotrichosis.

Johnson square procedure for lentigo maligna and lentigo maligna melanoma.

BACKGROUND: Lentigo maligna (LM) and lentigo maligna melanoma (LMM) can be difficult to manage surgically. Predetermined margins can be inadequate because of subclinical spread, or can affect function when margins are adjacent to the eye or mouth. AIM: To describe our 5-year experience in Nottingham of using the staged square procedure (Johnson square) in excising difficult facial LM and LMM. METHODS: The square procedure is a staged technique useful for ill-defined lesions and for lesions that have a high recurrence rate due to subclinical spread. It uses paraffin wax-embedded peripheral vertical sections for margin control, ensuring complete clearance as the surgical margins are usually examined at distances of 2-5 mm from the periphery of the lesion. RESULTS: We treated 21 patients with LM or LMM with the staged square procedure over a 5-year period. Of the 21 patients, 10 needed only one stage of surgery, 6 needed two stages, 3 needed three stages and 2 needed four stages. To date, there has been only one recurrence, which was of an extensive lesion that crossed the medial canthus, making margin control impossible because of the anatomical limitations. CONCLUSIONS: The staged square procedure is an effective treatment for LM and LMM. It attempts to conserve tissue while ensuring a higher clearance rate. This offers favourable cosmetic outcomes and better prognosis, especially for facial LM and LMM.

Primary cutaneous B-cell lymphoma in Nottinghamshire U.K.: prognosis of subtypes defined in the WHO-EORTC classification.

BACKGROUND: Primary cutaneous B-cell lymphomas (PCBCL), with the exception of large B-cell lymphoma of leg type and intravascular large B-cell lymphoma, are associated with an excellent prognosis. These lymphomas have become much better understood in recent years leading to the publication in 2005 of the World Health Organization-European Organisation for Research and Treatment of Cancer classification. OBJECTIVES: To determine the relative frequency of occurrence of subtypes of PCBCL in a defined population, and the survival of patients with these subtypes. METHODS: During the period 1987-2009, 61 consecutive patients with PCBCL were identified from the Nottingham Lymphoma Registry (population 1·1 million). After histological review, the number of patients with each subtype was as follows: marginal zone, 18; follicle centre, 14; diffuse large B cell, leg type, 16; diffuse large B cell, other sites, 12; and intravascular large B cell, one. RESULTS: The 5- and 10-year lymphoma-specific survival for patients with marginal zone lymphoma was 100%. The only patient with intravascular large B-cell lymphoma died from widespread disease in spite of chemotherapy. The 4-year lymphoma-specific survival for follicle centre cell lymphoma was 90%. Patients with the other subtypes had the following 5-year lymphoma-specific survival rates: diffuse large B cell, leg type, 61% and diffuse large B cell, other, 40%. The median age at diagnosis for patients with diffuse large B-cell lymphoma, leg type was 82 years and as a consequence the 5-year overall survival was only 15%. There was a 3·4-fold increase in the incidence of PCBCL from the period 1987-1997 to the period 1998-2009. CONCLUSIONS: PCBCL is a rare disease (incidence around three per million population per year). It is, in our view, essential that it is diagnosed by a pathologist with an interest in cutaneous lymphoma and that the very different prognosis of the individual subtypes is appreciated by the treating clinician.

Anti-neutrophil cytoplasmic antibody-positive neutrophilic dermatosis of the dorsal hands.

Neutrophilic dermatosis of the hands is a localized variant of Sweet syndrome (SS). It was first reported in 1995, and is an uncommon condition, with < 100 cases reported to date. The female preponderance, morphological and histological features, and response to treatment are similar to SS, but it differs in its distribution on the body. There may also be a lack of systemic features and inconsistent laboratory findings. Significantly, about half of all cases are associated with haematological problems, i.e. myelodysplasia and leukaemia. Other cases may be associated with ulcerative colitis or solid tumours. We describe a case of a 71-year-old man with neutrophilic dermatoses of the hands, who also had involvement of the lips. There was an associated rise in his anti-neutrophil cytoplasmic antibody level, which corresponded with the activity of the disease.

A novel case of follicular mucinosis after autologous stem-cell transplantation for multiple myeloma.

Follicular mucinosis is a rare inflammatory disorder of unknown aetiology, characterized by mucin deposition in hair follicles and sebaceous glands. FM can occur as a benign idiopathic primary disorder or secondary to malignant lymphoproliferative processes, most notably mycosis fungoides. We report a novel case of FM developing after autologous stem-cell transplantation for multiple myeloma, a correlation not previously reported in the literature.

An ultrastructural study of lymphocytes with surface-bound immunoglobulin.

This report is on a radioautographic study of lymphocytes exposed to (125)I-labeled anti-Ig in an attempt to identify surface-bound Ig molecules. The results as studied by ultrastructural radioautography confirmed the presence of surface-bound Ig on a certain population of lymphocytes. The specificity of the anti-Ig was determined by using appropriate controls that included the use of an absorbed anti-Ig and anti-hemocyanin antibody. The labeling pattern resulting from the interaction of labeled anti-Ig and Ig was found to be specifically associated with the cell surface and random in its distribution. Morphological differences were not apparent between labeled and nonlabeled lymphocytes in the spleen and lymph nodes. In the thymus, most lymphocytes did not exhibit detectable Ig. The few thymic lymphocytes that were labeled had unique morphological characteristics that included fewer ribosomes, many of which were monoribosomes. Relative to the amount in their cytoplasmic organelles, plasma cells had surface Ig but to a lesser degree than lymphocytes. Finally, macrophages were nonspecifically labeled and contained antibody on their membranes as well as intracellularly.

Ligand-induced movement of lymphocyte membrane macromolecules. I. Analysis by immunofluorescence and ultrastructural radioautography.

The fate of different complexes on the membrane of thymocytes and spleen lymphocytes was studied with the use of both immunofluorescence and ultrastructural radioautography. The complexes of anti-immunoglobulin (Ig) with the surface Ig of B lymphocytes were present all around the membrane at 4 degrees C; an increase in temperature produced a rapid aggregation of the complex into a cap which was readily interiorized in vesicles. Ultrastructural details of this process were given. The movement of the complexes depended upon the amount of anti-Ig and the temperature. The complexes of anti-lymphocyte antibody with surface antigen(s) did not result in formation of a single large aggregate (or cap) unless an anti-antibody was brought into the reaction. The caps formed by this trilayered complex were not interiorized. Concanavalin A (Con A) bound to cell surface carbohydrate moieties and the complexes of Con A readily formed a cap and were interiorized. Finally, antibodies to H-2 determinants did not form in most instances a single cap aggregate even when anti-antibodies were used. With time the H-2 complexes tended to form several large aggregates with some endocytosis.

Chemical inhibitors of phagosome-lysosome fusion in cultured macrophages also inhibit saltatory lysosomal movements. A combined microscopic and computer study.

The effects on lysosomal movements produced by the weak base ammonium chloride and by a representative polyanion poly-D-glutamic acid (PGA), previously reported to inhibit phagosome-lysosome (P-L) fusion, have been studied in cultured mouse macrophages using direct visual phase-contrast microscopy, a previously described (1, 3, 7) fluorescence assay of fusion, and computer analysis techniques. Treatment of the macrophages with 5-10 mM NH4Cl for 0.5-2 h or with 100 micrograms PGA/ml for 5 d caused a striking inhibition of saltatory lysosomal movements, as well as the expected inhibition of P-L fusion. Two other anionic fusion inhibitors tested, dextran sulphate and suramin, inhibited movements similarly. Removal of the NH4Cl from the cell medium reversed the lysosomal stasis and restored P-L fusion. Computer analyses of changes in lysosomal positions in treated and untreated macrophages during 2, 10, and 30-s intervals, using data from photomicrographs, computer graphics, and quantitative nearest-neighbour techniques developed for this purpose, supported the qualitative visual observation of the inhibition of lysosomal movements by the fusion inhibitors NH4Cl and PGA. Over the chosen intervals, from 80 to 96% of the lysosomes could be paired within 1 micron of each other in the NH4Cl- and PGA-treated cells in comparison with 50-70% in normal cells. The differences between the drug-treated and normal cells were highly significant. In an analogous system, the lysosomal stasis induced by hypertonic sucrose was examined and it was observed that P-L fusion too was inhibited. Both effects were reversible. We conclude that inhibition of P-L fusion and of lysosomal movement are associated. We suggest a causal relationship between these changes, namely, that the lysosomotropic inhibitors of fusion under study produce their effects largely, though perhaps not exclusively, by reducing saltatory lysosomal motion and consequently periphagosomal assembly, rather than directly and independently on P-L contact or on the fusion process itself. The possibility is raised that microtubules may be involved in the effector mechanism of these modulations.

Immunoglobulin-positive cells: their appearance in the mixed lymphocyte reaction.

In mixed lymphocyte cultures prepared with thoracic duct lymphocytes from allogeneic rats, approximately 20 percent of all blast cells that appeared at the end of 72 hours of incubation had surface receptors for rabbit antibody to rat immunoglobulin.

Role of lipid polymorphism in pulmonary surfactant.

The development of artificial surfactants for the treatment of respiratory distress syndrome (RDS) requires lipid systems that can spread rapidly from solution to the air-water interface. Because hydration-repulsion forces stabilize liposomal bilayers and oppose spreading, liposome systems that undergo geometric rearrangement from the bilayer (lamellar) phase to the hexagonal II (HII) phase could hasten lipid transfer to the air-water interface through unstable transition intermediates. A liposome system containing dipalmitoylphosphatidylcholine was designed; the system is stable at 23 degrees C but undergoes transformation to the HII phase as the temperature increases to 37 degrees C. The spreading of lipid from this system to the air-water interface was rapid at 37 degrees C but slow at 23 degrees C. When tested in vivo in a neonatal rabbit model, such systems elicited an onset of action equal to that of native human surfactant. These findings suggest that lipid polymorphic phase behavior may have a crucial role in the effective functioning of pulmonary surfactant.

Biofilm penetration, triggered release and in vivo activity of inhaled liposomal amikacin in chronic Pseudomonas aeruginosa lung infections.

OBJECTIVES: Chronic infections of Pseudomonas aeruginosa in the lungs of cystic fibrosis patients are intractable antibiotic targets because of their biofilm mode of growth. We have investigated the biofilm penetration, mechanism of drug release and in vivo antimicrobial activity of a unique nanoscale liposomal formulation of amikacin designed specifically for nebulization and inhaled delivery. METHODS: Penetration of fluorescently labelled liposomes into sputum or P. aeruginosa (PA3064) biofilms was monitored by a filter assay and by epifluorescence or confocal scanning laser microscopy. Amikacin release in vitro and rat lung levels after inhalation of nebulized material were measured by fluorescence polarization immunoassay. A 14 day agar bead model of chronic Pseudomonas lung infection in rats was used to assess the efficacy of liposomal amikacin versus free aminoglycosides in the reduction of bacterial count. RESULTS: Fluorescent liposomes penetrated readily into biofilms and infected mucus, whereas larger (1 microm) fluorescent beads did not. Amikacin release from liposomes was mediated by sputum or Pseudomonas biofilm supernatants. Rhamnolipids were implicated as the major releasing factors in these supernatants, active at one rhamnolipid per several hundred lipids within the liposomes. Inhaled liposomal amikacin was released in a slow, sustained manner in normal rat lungs and was orders of magnitude more efficacious than inhaled free amikacin in infected lungs. CONCLUSIONS: Penetration of biofilm and targeted, sustained release from liposomes can explain the superior in vivo efficacy of inhaled liposomal amikacin versus free drug observed in a 14 day infection model. Inhaled liposomal amikacin may represent an important therapy for chronic lung infections.

Interventions for basal cell carcinoma of the skin.

BACKGROUND: Basal cell carcinoma (BCC) is the commonest skin cancer. BCCs are slow-growing, locally invasive, epidermal skin tumours which mainly affect white skinned people. The first line treatment is usually surgical excision, but numerous alternatives are available. OBJECTIVES: To assess the effects of treatments for basal cell carcinoma. SEARCH STRATEGY: We searched the Cochrane Skin Group Specialised Register (January 2006), the Cochrane Central Register of Controlled Trials (The Cochrane LIbrary Issue 1, 2006), the Cochrane Database of Systematic Reviews (The Cochrane Library Issue 1, 2006), MEDLINE (2004 to January 2006), EMBASE (2005 to January 2006), the metaRegister of Controlled Trials (February 2006). Cited references of all trials identified and key review articles were searched. Pharmaceutical companies were contacted where appropriate for reviews or unpublished trials. SELECTION CRITERIA: Inclusion criteria were adults with one or more histologically proven, primary basal cell carcinoma. The primary outcome measure was recurrence at three to five years, measured clinically. The secondary outcome included early treatment failure within six months, measured histologically. Adverse treatment effects included aesthetic appearance and pain during and after treatment. DATA COLLECTION AND ANALYSIS: Two authors independantly carried out study selection and assessment of methodological quality. MAIN RESULTS: Twenty seven studies were identified. Only one RCT of surgery versus radiotherapy had primary outcome data at four years, showing significantly more persistent tumours and recurrences in the radiotherapy group as compared to the surgery group, (RR 0.09, 95%CI, 0.01 to 0.69). One study found no significant difference for recurrence at 30 months when Moh's micrographic surgery was compared to surgery for high risk facial BCCs, (RR 0.64, 95%CI 0.16,2.64). One study of methylaminolevulinate photodynamic therapy (MAL PDT) versus cryotherapy found no significant difference in recurrences in the MAL PDT group when compared to cryotherapy at one year (RR 0.50, 95% CI 0.22,1.12). Cryotherapy showed no significant difference in recurrences at one year when compared to surgery on one small study. When radiotherapy was compared to cryotherapy there were significantly fewer recurrences at one year in the radiotherapy group compared to the cryotherapy group.Short-term studies suggest a success rate of 87 to 88% for imiquimod in the treatment of superficial BCC using a once-daily regimen for 6 weeks and a 76% treatment response when treating nodular BCC for 12 weeks, when measured histologically. AUTHORS' CONCLUSIONS: Overall there has been very little good quality research on treatments for BCC. Most trials have only evaluated BCCs in low risk locations. Surgery and radiotherapy appear to be the most effective treatments with surgery showing the lowest failure rates. Although cosmetic outcomes appear good with PDT, long term follow up data are needed. Other treatments might have some use but few have been compared to surgery. An ongoing study comparing imiquimod to surgery should clarify whether imiquimod is a useful option.

Interventions for preventing non-melanoma skin cancers in high-risk groups.

BACKGROUND: Some groups of people have a greater risk of developing common non-melanoma skin cancers (NMSC). OBJECTIVES: To evaluate interventions for preventing NMSC in people at high risk of developing NMSC. SEARCH STRATEGY: We searched the Cochrane Skin Group Specialised Register (March 2007), the Cochrane Central Register of Controlled Trials (The Cochrane Library Issue 1, 2007, MEDLINE (from 2003 to March 2007), EMBASE (from 2005 to March 2007), the metaRegister of Controlled Trials (February 2007). References from trials and reviews were also searched. Pharmaceutical companies were contacted for unpublished trials. SELECTION CRITERIA: Randomised controlled trials of adults and children at high risk of developing NMSC. DATA COLLECTION AND ANALYSIS: Two review authors independently selected studies and assessed their methodological quality. MAIN RESULTS: We identified 10 trials (7,229 participants) that assessed a variety of interventions. One trial found T4N5 liposome lotion significantly reduced the rate of appearance of new BCCs in people with xeroderma pigmentosum. One of three trials of renal transplant recipients showed a significantly reduced risk of new NMSCs when acitretin was compared to placebo (relative risk (RR) 0.22 95% confidence interval (CI) 0.06 to 0.90) and no significant difference in risk of adverse events in two trials (RR 1.80, 95% CI 0.70 to 4.61). In three trials conducted in people with a history of NMSC, the evidence was inconclusive for the development of BCCs for retinol or isoretinoin. However the risk of a new SCC in one trial (HR 1.79, 95% CI 1.16 to 2.76) and adverse events in another trial (RR 1.76 95% CI 1.57 to 1.97) were significantly increased in the isotretinoin group compared with placebo. In one trial selenium showed a reduced risk of other types of cancer compared with placebo (RR 0.65, 95% CI 0.50 to 0.85) but also a significantly elevated risk of a new NMSC (HR 1.17 95% CI 1.02 to 1.34). The evidence for one trial of beta-carotene was inconclusive; and there was a trend towards fewer new NMSC in a trial of a reduced fat diet (RR 0.16, 95% CI 0.02 to 1.31), p=0.09. AUTHORS' CONCLUSIONS: Some preventative treatments may benefit people at high risk of developing NMSC, but the ability to draw firm conclusions is limited by small numbers of trials, often with one trial per intervention or with inconsistent results between studies.

Optimisation of slow-pyrolysis process conditions to maximise char yield and heavy metal adsorption of biochar produced from different feedstocks.

The objective of this work was to identify biomass feedstocks and optimum pyrolysis process conditions to produce a biochar capable of adsorbing metals from polluted groundwater. Taguchi experimental design was used to determine the effects of slow-pyrolysis process conditions on char yield and zinc adsorption. Treatments were repeated using six candidate feedstocks (Lolium perenne, Lolium perenne fibre, Miscanthus x giganteus, Salix viminalis, Fraxinus excelsior and Picea sitchensis) and the resultant chars were tested for metal adsorption performance. Chars produced from L. perenne and its extracted fibre displayed the greatest zinc adsorption performance and removed 83.27-92.96% respectively. Optimum process conditions in terms of both char yield and zinc adsorption performance were achieved from slow-pyrolysis at 300°C for 2h using a feedstock with a particle size of less than 1mm.

The captured volume of multilamellar vesicles.

Characterization of classical 'hand-shaken' multilamellar lipid vesicles (MLVs) confirmed that these systems exclude solute during formation thus confounding previous captured volume measurements which typically have utilized solute as a merker of the occluded aqueous space. We used solvent rather than solute to determine the captured volume of these systems and obtained values at least twice those previously reported. We present here a captured volume and lamellarity profile of 'hand-shaken' MLVs and suggest that these parameters are dependent on the lipid concentration present during hydration.

Liposomal entrapment of the neutrophil-derived peptide indolicidin endows it with in vivo antifungal activity.

Indolicidin, a cationic tridecapeptide amide isolated from the granules of bovine neutrophils, has been found to possess potent antimicrobial activity in vitro but its nonselective toxicity could restrict its therapeutic utility. We found that the concentration at which indolicidin disrupts washed human red blood cell membranes coincided with the concentration at which indolicidin self associates. Because of a preponderance of hydrophobic residues, we believed that indolicidin would partition into liposomes which would restrict its exchange with biological tissues and consequently reduce its toxicity. Fluorescence spectroscopy of indolicidin added to 100 nm liposomes comprised of POPC, POPC/cholesterol (60:40 mol%), DPPC, or DPPC/cholesterol (60:40) revealed a large blue-shift and an increase in intensity of the emission profile indicating insertion into the bilayer. Of the lipids tested, POPC exhibited the highest degree of indolicidin binding as determined by fluorescence and encapsulation efficiency. By sequestering indolicidin within the lipid bilayer of 100 nm POPC liposomes we significantly reduced its toxicity to CHO/K1 cells. Likewise, the systemic toxicity of liposomal indolicidin in Balb/c mice was decreased dramatically relative to aqueous solutions; the maximum dose at which no deaths occurred was 0.4 mg/kg for free indolicidin versus 40 mg/kg for indolicidin-POPC. Because of this decrease in toxicity, we were able to administer liposomally encapsulated material at significantly higher concentrations than unencapsulated aqueous material and achieve efficacy in treating animals systemically infected with Aspergillus fumigatus. Liposomal but not free indolicidin was found to be effective in obtaining cures. This report is the first description of the in vivo therapeutic activity of a neutrophil-derived antimicrobial peptide and suggests that liposomal treatment modalities will provide effective strategies for endowing this class of compounds with pharmacological utility.

Curvature dependent induction of the interdigitated gel phase in DPPC vesicles.

Ethanol causes biphasic melting behavior in saturated lecithins (Rowe (1983) Biochemistry 22, 3299-3305), a consequence of the formation of the stable interdigitated phase (Simon, S.A. and McIntosh, T.J. (1984) Biochim. Biophys. Acta 773, 169-172). The membrane systems studied to date have been large vesicle systems in which the membrane surface can be assumed to be locally planar. An immediate question arises as to whether surfaces of higher curvature interdigitate. To address this question we have prepared DPPC vesicles of varying diameters which we employed to determine the limiting size at which interdigitation occurs using ethanol as the inducer. We find that with decreasing vesicle size the concentration of ethanol necessary for the onset of interdigitation increases. Small isolated vesicles, at inducing concentrations of ethanol, do not stably interdigitate but rupture and coalesce into a viscous gel comprised of interdigitated lipid sheets. As discussed elsewhere (Ahl et al. (1992) Biophys. J. 243a) these sheets can be used as precursors for producing liposomes of large size and high internal volumes useful in drug delivery or modeling applications.